Expression of Mx protein in tissues of Atlantic salmon post-smolts--an immunohistochemical study.

Abstract

A rabbit antiserum was produced from a 12-amino acid long peptide common to the 3 known isoforms of Atlantic salmon Mx proteins. The antibody stained ASK-1 cells 48h after stimulation with poly I:C. In Western blots of these cells, the antibody stained a doublet with MW about 75kDa and another band at about 65kDa, typical of the MW of Atlantic salmon Mx. Western blots of kidney from IPNV-injected salmon showed a similar staining pattern. In immunohistochemistry, the antibody stained the gill, kidney and liver tissue of a fish infected with IPNV by cohabitation. These tissues also expressed high levels of interferon (IFN) and Mx transcripts as determined by real-time qRT-PCR. Normal healthy salmon post-smolts sampled at 4-8 weeks after transfer to sea water had very low-level expression of IFN and Mx transcripts. However, at 4 and 5 weeks after sea water transfer the gill, kidney and liver of these fish stained strongly for Mx protein. Thereafter, immunostaining of Mx markedly diminished in all tissues, persisting weakly in the gill. It has been reported that Atlantic salmon smolts constitutively express IFN and Mx transcripts around the time of smolting. Presumably the Mx protein detected in the tissues for about 6 weeks after transfer to sea water resulted from such a transcriptional event. As Mx is known to provide protection against IPNV infections it is tempting to associate the duration of persistence of Mx protein with the outbreaks of IPN-related mortalities in post-smolts, 6-8 weeks after transfer to sea water.